Adeno-associated virus (AAV) packaging limits constrain the design, performance, and in vivo application of genetically encoded biosensors. We developed a split intein-mediated reconstitution strategy enabling modular delivery and reassembly of oversized fluorescence lifetime-based biosensors. Using this platform, we engineered an oversized cAMP sensor compatible with one-photon fluorescence lifetime measurements, enabling monitoring of intracellular signaling dynamics in distinct neuronal subtypes in freely behaving mice.
Lee, J. O., Puhl, H., Holder, A., Sheridan, A., Darden, C., Shah, M., Nguyen, T., Hines, K., Augustin, S. M., Kim, Y., Vogel, S. S., Lovinger, D. M.
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